Made available in DSpace on 2014-06-12T23:13:55Z (GMT). No. of bitstreams: 2 arquivo9519_1.pdf: 1411821 bytes, checksum: a13d197e45c98cda482e7ab8a29fe326 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2012 / O leite humano é o alimento ideal para o lactente e nenhum outro alimento poderá substituí-lo com vantagem. A amamentação forma uma base biológica e emocional tanto para a saúde da mãe quanto para da criança. Apesar da excelência do leite humano, o desmame precoce é muito freqüente no Brasil, principalmente na região Nordeste. A mastite puerperal ou da lactação é um processo inflamatório das glândulas mamárias que acomete mulheres em fase de lactação, que devido ao desconforto e à dor torna-se importante causa de desmame precoce e quando não tratada corretamente pode evoluir para abscessos. Os principais agentes etiológicos das mastites puerperais são os Staphylococcus aureus (S. aureus, micro-organismos importantes e que vêm apresentando resistência a antibióticos administrados na clínica médica. Diante destes aspectos, este estudo teve como objetivo Investigar a prevalência da mastite de lactação e dos principais agentes etiológicos a fim de fortalecer o estímulo à amamentação no município de Vitória de Santo Antão, no período de agosto de 2010 a março de 2011 a fim de subsidiar os profissionais de saúde à prevenção, detecção e intervenção precoce das mastites.Trata-se de um estudo descritivo do tipo transversal e para concretização do estudo foram coletadas e avaliadas amostras de leite em frasco estéril, amostras da microbiota da pele das mãos das lactantes através do toque das polpas digitais em placas de Agar Manitol Sal e fossas nasais de lactentes e lactantes através de swabs em 67 mulheres de zona rural e urbana e seus respectivos filhos. Utilizou-se os testes de qui-quadrado e teste Exato de Fisher pelo SPSS na versão 13.0 para avaliação das variáveis. Todas as amostras foram devidamente encaminhadas ao Laboratório de Microbiologia dos Alimentos do Centro Acadêmico de Vitória. Para isolamento e identificação de S. aureus, foi realizado o teste de coloração pelo método de Gram e testes bioquímicos como os de catalase, coagulase, DNase e fermentação de glicose e manitol em aerobiose e anaerobiose. Realizou-se o testes de sensibilidade antimicrobiana (TSA) in vitro para as cepas de S.aureus, pelo método de disco-difusão, utilizou-se os antibióticos disponíveis na rede de saúde do SUS para uso em lactantes. Foi aplicado um questionário estruturado com questões fechadas com intuito de conhecer os principais fatores de risco associados às mastites e obter informações completares para a pesquisa. O estudo mostrou uma prevalência de 17,9% de mastite, tendo como principal agente etiológico Staphylococcus Coagulase Negativa SCN com 50,8%. Em 8,3% das mastites não houve crescimento microbiano. Com 16,7% o S.aureus representou o terceiro micro-organismo mais prevalente nas mastites. Quanto as amostras de mão e fossas nasais o SCN apresentou maior freqüência seguido de S.aureus. O S.aureus apresentou maior resistência a Penicilina G com 93,9% e maior sensibilidade a amicacina com 96,3%. Foi comprovado que a dor no mamilo, ingurgitamento mamário e fissura mamilar são aspectos relacionados com a mastite puerperal. Ao fim do estudo foi elaborado um instrumento de informação sobre casos de mastite para acompanhamento da incidência e elaboração de políticas de prevenção a doença

Submitted by Nayara Passos (nayara.passos@ufpe.br) on 2015-03-04T13:23:34Z No. of bitstreams: 2 2012-Dissertação-Hebe-Duarte.pdf: 1411800 bytes, checksum: 25c5fd3543df1c83001eb97f6172b7ac (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-03-04T13:23:34Z (GMT). No. of bitstreams: 2 2012-Dissertação-Hebe-Duarte.pdf: 1411800 bytes, checksum: 25c5fd3543df1c83001eb97f6172b7ac (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2012 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Staphylococcus aureus is an important opportunistic pathogen in humans and animals. In humans, there has been an increase in community-associated methicillin-resistant Staphylococcus aureus (MRSA) causing disease in healthy humans. The exact evolutionary origins and basis for its recent expansion are not yet clear. In livestock, S. aureus is an important cause of diseases of welfare and economic importance, including bovine mastitis. Molecular typing studies demonstrate that natural populations of S. aureus are highly clonal and largely adapted to a specific host, however there are some lineages that colonise multiple host species. In particular, clonal complex 97 (CC97) is a dominant bovine mastitis-associated lineage which has been isolated from other animal species, and more recently there are increasing reports of CC97 S. aureus from human infections worldwide. The basis for this wide host tropism is currently unknown. In order to investigate the evolutionary origins of S. aureus CC97, 43 strains were selected for whole genome sequencing, isolated from humans, cattle and pigs, from 18 different countries on 4 continents, ranging from 1956 to 2012. Phylogenetic analysis using high quality core genome single nucleotide polymorphisms (SNPs) resolved the single CC97 lineage into host-adapted sublineages, which were likely the result of 2 independent livestock-to-human host jumps estimated to have occurred at least 40 years ago. One of the human sublineages consisted of strains from 4 continents indicating global dissemination since the host jump occurred. In order to investigate the genetic basis for human host adaptation in S. aureus CC97, comparative genomic analysis of mobile genetic elements, nonsynonymous SNPs and small insertions and deletions was performed. Of note, independent acquisitions of genetic elements encoding antimicrobial resistance and specific mediators of human innate immune evasion were identified in the human-adapted S. aureus CC97 strains. These data are consistent with an important role for mobile genetic elements in the host adaptive evolution of S. aureus CC97. Also in the current study, a bovine-associated single locus variant of ST97 (ST71) was identified as a phylogenetic outgroup relative to all other S. aureus CC97 strains examined. Comparative genomic analysis of ST71 strains with representative bovine ST97 strains indicate that ST71 has a mosaic genome. A large region spanning the origin of replication demonstrated closest homology to non-CC97 ruminant-associated genotypes, with the remainder of the genome consistent with an ancestral ST97 genetic background. Recombination detection analysis predicts that one or more large-scale recombination events have occurred in the region that spans the origin of replication, resulting in variation in gene content between ST71 and ST97. The data highlight the potential role of homologous recombination in rapidly generating genomic diversity that might alter the phenotype of strains in the ecological niche of the bovine mammary gland. Overall, the study reveals the evolutionary history of a major pathogenic clone of S. aureus affecting multiple host species, and identifies the genetic events which have contributed to its success.

Memoria para optar al título de Ingeniero en Alimentos / La desinfección de verduras y frutas es un tratamiento, destinado a reducir la microflora presente en forma natural en estos alimentos o aquella que se incorpora a través de las distintas etapas que ocurren desde su cultivo, hasta que el producto sea expendido y consumido. La desinfección también abarca las superficies y equipos de trabajo, donde se procesan y manipulan alimentos, para así asegurar la inocuidad al consumidor. Se realizó un estudio comparativo in vitro de la actividad germicida de un desinfectante, compuesto por una mezcla de monoéster de propilenglicol, ácido láctico y ácido cítrico, entre otros. Este fue probado a diferentes tiempos de acción y concentraciones, incluyendo la recomendada por el fabricante. Las pruebas se realizaron frente a cepas de Staphylococcus aureus salvaje y ATCC 29213 y Escherichia coli salvaje y ATCC 25922. Se determinó la cinética de muerte de los microorganismos para así evaluar la eficacia germicida del desinfectante, la velocidad específica de muerte, coeficiente de dilución y el tiempo de reducción de los microorganismos. La eficiencia germicida in vitro del desinfectante a 400 ppm fue de 99,999%, para Escherichia coli salvaje, Escherichia coli ATCC 25922 y Staphylococcus aureus ATCC 29213 a los 15 minutos, lo cual demuestra que es un buen bactericida, para dichas cepas. Sin embargo, para Staphylococcus aureus salvaje solo se observó un 99,850% de eficacia a la misma concentración y tiempo. Staphylococcus aureus salvaje y ATCC 29213 resultó más resistente que Escherichia coli salvaje y ATCC 25922 lo cual se demuestra por los valores obtenidos para el tiempo de reducción decimal y velocidad específica de muerte / The disinfection of vegetables and fruits is a treatment aimed to reduce the microflora that grows naturally in these foodstuffs or that is incorporated through different occurring stages from cultivation until the product is sold and consumed. Disinfection also must be carried out on the working surfaces and equipments where food is processed and manipulated, to ensure the safety for the consumer. In this study, an in vitro comparative study of the germicide activity of a disinfectant composed of a mixture of monoester of propylene glycol, lactic acid and citric acid among others, was performed. This product was tested at different action times and concentrations, including those recommended by the manufacturer. The tests were carried out against wild type and ATCC strains of Staphylococcus aureus (ATCC 29213) and Escherichia coli (ATCC 25922). The evaluated parameters were the kinetics of death of the microorganisms, the germicide efficacy of the disinfectant, the specific death rate, the dilution coefficient and the reduction time of the microorganisms. The in vitro germicide efficiency of the disinfectant at 400 ppm during 15 minutes of contact was 99,999%, for the wild type of Escherichia coli, for Escherichia coli ATCC 25922 and for Staphylococcus aureus ATCC 29213. This fact demonstrates that the disinfectant has a good bactericide effect on the assayed microorganisms; however, for the wild type of Staphylococcus aureus only a 99, 850% of efficacy using the same concentration and time was observed. The wild type of Staphylococcus aureus and the ATCC 29213 strain showed a greater resistance than the two strains of Escherichia coli, which is demonstrated in the results obtained for the decimal reduction time and specific death rate

Class of 2013 Abstract / Specific Aims: The purpose of this study was to assess risk factors that are associated with inpatient charges and mortality rates with methicilin-resistant Staphylococcus aureu (MRSA), Aspergillus, and Candida in children with leukemia. The hypotheses is MRSA and opportunistic fungal infections are associated with higher inpatient charges and worse outcomes relative to those without. Children with leukemia are especially at risk due to underdeveloped and compromised immune systems. The rational is that identifying the risk factors that affect mortality and inpatient charges of these infections should add to the current knowledge of treating and preventing these infections in immune compromised patients. Methods: Retrospective cohort study using the Agency for Healthcare Research Quality (AHRQ) KIDS 2009 database. Inclusion criteria was defined as 17 years of age or younger and a ICD-9 code for an active infection with MRSA, Aspergillus, or Candida. Regression analysis's were performed to identify factors that had an impact on mortality, length of stay, and direct patient charges. Main Results: Values that were significant (p≤0.05) for predicting an increase in mortality were Age (year), MRSA, Aspergillus, Candida, and Deyo-Charlson comorbidity scores. Significant predictors of increased Inpatient Charges (p≤0.05) was Age (year), MRSA, Aspergillus, Candida, Female Sex, Deyo-Charlson comorbidity scores, Urban Hospitals, and the Southern Region. Significant predictors of Increased Length of Stay (p≤0.05) MRSA, Aspergillus, Candida, Female Sex, Deyo-Charlson, Urban Hospitals, Teaching Hospitals, and the Southern Region. Conclusion: Although uncommon in leukemia cases involving pediatrics or young adults, statistically significant and large risks of higher mortality, length of stay, and inpatient charges were noted in cases involving MRSA, aspergillus, and candida. Increasing Deyo-Charlson comorbidities scores were also consistently important predictors for poor outcomes in these leukemia patients and, with certain outcomes, increasing age and female sex.

Reporter gene technology has facilitated greatly the analysis of gene expression and the study of individual promoters and their regulation. Although various reporter gene systems are available, none of them are universally applicable and consequently, studies aimed at screening of new reporters are continuing. Toward this end, an acid phosphatase, designated SapS, was identified and characterized from the culture supernatant of a Staphylococcus aureus strain isolated from vegetables. Biochemical characterization of the 30-kDa monomeric enzyme indicated that it displayed optimum activity at 40°C and pH 5, using p-nitrophenyl phosphate (pNPP) as substrate. The enzymatic activity was enhanced by Mg2+, but was inhibited by EDTA and molybdate. Based on its properties and amino acid sequence analyses, SapS was classified as a new member of the bacterial class C family of non-specific acid phosphatases. The S. aureus SapS enzyme was subsequently evaluated as a reporter for host strain evaluation and cell surface display. Bacillus halodurans of which the major cell wall protease gene (wprA) was inactivated was used as expression host, and the cell wall-binding domain of the cwlC gene from B. halodurans was used as an anchoring motif for cell surface display. The results from in vitro enzyme activity assays indicated that extracellular production of the SapS reporter enzyme was improved 3.5-fold in the mutant compared to wild-type B. halodurans strain. Zymographic detection of SapS activity showed that the SapS-CwlC fusion protein was localized in the B. halodurans cell wall fraction, thus demonstrating the potential of SapS as a reporter for cell surface display of heterologous proteins. The versatility of the SapS enzyme as a reporter for gene expression and protein secretion in both Gram-positive and Gram-negative bacteria was also investigated. Transcriptional and translational fusions of the sapS gene with selected heterologous promoters and signal sequences were constructed, and expressed in Escherichia coli, B. subtilis and B. halodurans. The strongest promoter for heterologous protein production in each of the host strains was identified, i.e. the E. coli lacZ promoter in E. coli, the B. halodurans alkaline protease promoter in B. subtilis, and the B. halodurans σD promoter in B. halodurans. / Thesis (PhD)--University of Pretoria, 2010. / Microbiology and Plant Pathology / unrestricted

Le genre Staphylococcus, première étiologie des infections ostéo-articulaires (IOA), est associé à des formes particulièrement difficiles à traiter. Trois mécanismes phénotypiques ont été rattachés à ce fort taux de chronicité et de rechutes, permettant l'adaptation bactérienne à la vie au sein du tissu osseux et un échappement au système immunitaire de l'hôte et à l'action des antibiotiques : la formation de biofilm, la persistance des staphylocoques dans les ostéoblastes, et l'évolution vers le morphotype de small colony variant (SCV). Longtemps considéré comme simple commensal cutanéo-muqueux, S. epidermidis est désormais reconnu comme un agent étiologique majeur des IOA sur matériel. Or, si le portage est universel, l'infection est un phénomène rare. A ce jour, aucun facteur génotypique n'a pu être associé au pouvoir invasif de certaines souches de portage. Notre travail a permis de montrer l'absence de pouvoir discriminant des capacités d'internalisation des ostéoblastes et de formation de biofilm entre souches commensales et invasives. Par ailleurs, un très faible taux d'internalisation de S. epidermidis dans les ostéoblastes a été mis en évidence, suggérant une importance moindre de ce mécanisme dans la physiopathologie des IOA à S. epidermidis par rapport aux IOA à S. aureus. Les principales études ayant porté sur les capacités d'interaction de S. aureus avec les ostéoblastes et de formation de biofilm ont cherché à en explorer les mécanismes à partir de souches de laboratoire ou de souches représentatives de quelques clones de S. aureus résistants à la méticilline (SARM). Dans notre cas, nous avons souhaité étudier une large collection de souches cliniques de S. aureus (n=95) sensible à la méticilline (SASM) responsables d'IOA aiguës ou chroniques. La caractérisation des fonds génétiques de cette collection, puis en élargissant notre étude à des collections de différents villes françaises, a d'abord permis de décrire une forte prévalence du clone émergent de SASM CC398 dans les IOA en France / Pas de résumé en anglais

Submitted by Amauri Alves (amauri.alves@ufv.br) on 2015-11-03T16:31:04Z No. of bitstreams: 1 texto completo.pdf: 761509 bytes, checksum: e43a21fc075340daf4e14f9f668c7d07 (MD5) / Made available in DSpace on 2015-11-03T16:31:04Z (GMT). No. of bitstreams: 1 texto completo.pdf: 761509 bytes, checksum: e43a21fc075340daf4e14f9f668c7d07 (MD5) Previous issue date: 2015-02-24 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Staphylococcus aureus é um patógeno humano oportunista que apresenta riscos a saúde humana, é capaz de aderir em superfícies bióticas e abióticas e formar biofilmes, tornando as células mais protegidas e de difícil remoção. Células liberadas do biofilme podem se constituir em importante fonte de contaminação de alimentos, comprometendo a qualidade e a segurança dos mesmos. O objetivo deste estudo foi verificar o efeito da bacteriocina nisina e dos sanitizantes hipoclorito de sódio e ácido peracético, em concentrações subinibitórias, sobre a hidrofobicidade da superfície de poliestireno, sobre o crescimento e formação de biofilmes por estirpes de S. aureus e ainda, verificar a interferência da nisina sobre a composição e estrutura do biofilme desse patógeno. Células de Staphylococcus epidermidis ATCC 35984 foram usadas como referência para formação de biofilmes. O cultivo das bactérias foi feito em caldo Luria-Bertani (LB) ou em meio sintético (MS). A presença de genes de adesão foi determinada pela reação em cadeia da polimerase (PCR) e os três genes avaliados, icaA, icaD e clfB, foram encontrados nas estirpes COL e FRI 722 de S. aureus, enquanto a estirpe Embrapa 4018 de S. aureus e em S. epidermidis, apenas o gene icaA foi identificado. A hidrofobicidade da superfície de poliestireno foi avaliada por meio da medida do ângulo de contato e constatou-se que o meio LB reduziu a hidrofobicidade da superfície, dificultando a observação do efeito dos antimicrobianos sobre a mesma. O tratamento da superfície de poliestireno com MS adicionado de 1,34 mg/L de nisina reduziu a adesão de S. aureus. Concentrações subinibitórias de 2,01 mg/L; 1.500 mg/L e 0,40 mg/L respectivamente dos antimicrobianos nisina, hipoclorito de sódio e do ácido peracético foram adicionadas isoladamente ou combinadas entre si ao caldo LB e constatou-se a diminuição da formação de biofilmes pela cultura mista de estirpes de S. aureus e por S. epidermidis em microplacas de poliestireno quando os antimicrobianos agiram isoladamente. A composição de polissacarídeos e DNA nos biofilmes de S. aureus e S. epidermidis foi alterada quando o cultivo ocorreu na presença de 2,01 mg/L de nisina. Entretanto, o conteúdo em proteínas nos biofilmes formados pela cultura viiimista das estirpes de S. aureus estudadas ou por S. epidermidis não foi alterado pela presença de nisina no meio de cultura em concentração subinibitória. A estrutura do biofilme de S. aureus e S. epidermidis foi avaliada por microscopia confocal a laser, confirmando os resultados quantitativos de que a presença de concentração subinibitória de nisina reduz a formação de biofilmes por estas espécies. Estes resultados demonstram que a investigação de produtos alternativos para auxiliar no controle e combate aos biofilmes é estratégia promissora além de contribuir com informações sobre a composição do biofilme de S. aureus. / Staphylococcus aureus is an opportunistic human pathogen that represents risks to the human health and is able to adhere and form biofilms on biotic and abiotic surfaces, being cells more protected and difficult to remove. Cells released from biofilms can be an important source of food contamination, compromising quality and safety. The aim of this work was to investigate the effect of the bacteriocin nisin and the sanitizers sodi- um hypochlorite and peracetic acid, in subinhibitory concentration, on the polystyrene surface hydrophobicity, on the growth and biofilm formation by S. aureus strains and also, verify the interference of nisin on the composition and structure of the pathogen biofilm. As a control of biofilm formation, Staphylococcus epidermidis ATCC 35984 was used. Bacteria were cultivated in Luria-Bertani (LB) broth or in synthetic medium (SM). The presence of adhesion genes was determined by polymerase chain reaction (PCR) and the three evaluated genes, icaA, icaD, and clfB, were found in S. aureus COL and FRI 722, while in the strain Embrapa 4018 and in S. epidermidis, only icaA gene was identified. The hydrophobicity of the polystyrene surface was evaluated by the con- tact angle measurement and it was found that the LB broth reduced the surface hydro- phobicity by itself, hindering the observation of the antimicrobials effect on it. Treat- ment of the polystyrene surface with SM supplemented with 1.34 mg/L of nisin reduced S.aureus adhesion. Subinhibitory concentrations of 2.01 mg/L; 1,500 mg/L and 0.40 mg/L of the antimicrobials nisin, sodium hypochlorite, and peracetic acid, respectively, were added separately or combined with each other in LB broth and the reduction of biofilm formed by S. aureus strains and S. epidermidis in polystyrene microtiter plates occurred when the antimicrobials acted alone. The content of polysaccharides and DNA in the biofilms of S. aureus and S. epidermidis was altered when cultivated in the pres- ence of 2.01 mg/L of nisin. However, the protein percentage in the biofilm formed by the culture mix composed by the three studied strains of S aureus or by S. epidermidis was not modified by the presence culture medium supplemented with subinhibitory concentration of nisin. Biofilm formation by S. aureus and S. epidermidis was also evaluated by confocal laser microscopy, confirming the quantitative results in which xsubinhibitory concentrations of nisin reduces biofilm by these species. These results demonstrate that the investigation of alternative products to help control and eradicate biofilms is a potential strategy and also provides information on the composition of S. aureus biofilms.

This study examines the prevalence of methicillin-resistant Staphylococcus aureus in midwestern swine herds and the workers in those herds.

No description available.